Mitochondrial and DNA damage in bovine somatic cell nuclear transfer embryos
نویسندگان
چکیده
The generation of reactive oxygen species (ROS) and subsequent mitochondrial and DNA damage in bovine somatic cell nuclear transfer (SCNT) embryos were examined. Bovine enucleated oocytes were electrofused with donor cells and then activated by a combination of Ca-ionophore and 6-dimethylaminopurine culture. The H2O2 and ˙OH radical levels, mitochondrial morphology and membrane potential (ΔΨ), and DNA fragmentation of SCNT and in vitro fertilized (IVF) embryos at the zygote stage were analyzed. The H2O2 (35.6 ± 1.1 pixels/embryo) and ˙OH radical levels (44.6 ± 1.2 pixels/embryo) of SCNT embryos were significantly higher than those of IVF embryos (19.2 ± 1.5 and 23.8 ± 1.8 pixels/embryo, respectively, p < 0.05). The mitochondria morphology of SCNT embryos was diffused within the cytoplasm. The ΔΨ of SCNT embryos was significantly lower (p < 0.05) than that of IVF embryos (0.95 ± 0.04 vs. 1.21 ± 0.06, red/green). Moreover, the comet tail length of SCNT embryos was longer than that of IVF embryos (515.5 ± 26.4 μm vs. 425.6 ± 25.0 μm, p < 0.05). These results indicate that mitochondrial and DNA damage increased in bovine SCNT embryos, which may have been induced by increased ROS levels.
منابع مشابه
I-12: Nuclear Reprogramming in Bovin Somatic Cell Nuclear Transfer
Somatic cell nuclear transfer (SCNT or cloning) returns a differentiated cell to a totipotent status; a process termed nuclear reprogramming. Reproductive cloning has potential applications in both agriculture and biomedicine, but is limited by low efficiency. To understand the deficiencies of nuclear reprogramming, our research has focused on both candidate genes and global gene expression pat...
متن کاملP-115: Melatonin Increases Developmental Rate of In Vitro Mouse Somatic Cell Nuclear
Background: The beneficial effect of supplementing culture medium with melatonin has been reported during in vitro embryo development of species such as mouse, bovine and porcine. However, the effect of melatonin on the mouse somatic cell nuclear transfer remained unknown. Materials and Methods: In this study, we assessed the effects of various concentrations of melatonin (10-6 to 10-12 M) on t...
متن کاملO-7: Improved In Vitro Development of Cloned Bovine Embryos Using S-Adenosylhomocysteine,A Non-Toxic Epigenetic
Background: Development of cloned bovine embryos. Materials and Methods: Oocytes collection,oocyte denudation, oocyte enucleation, nuclear transfer, electrofusion, reconstructed embryo activation, culture of reconstructed and IVF embryo,and treatment with SAH post fusion interval Treatment of reconstructed embryos with TSA for 12 hours after activation, preparation of somatic donor cells, donor...
متن کاملP-86: Production of Cloned Mice by Somaticm Cell Nuclear Transfer
Background: For several years, mammalian cloning by splitting an early embryo or nuclear transfer into oocytes method has been successfully performed. Cloning is now also possible using adult somatic cells. Although it has now been 15 years since the first cloned mammals were generated from somatic cells using nuclear transfer (NT), the success rate for producing live offspring by cloning is lo...
متن کاملO-18: Epigenetic Modification of Cloned Embryo Development; State of ART
Background: At the outset of the somatic cell nuclear transfer (SCNT) process, the chromatin structure of the somatic cell which governs its state of differentiation undergoes dramatic changes, called reprogramming, and is compelled back to the embryonic stage. However, the overall epigenetic makeup of the resultant cloned embryos has been acknowledged far different from the fertilized embryos....
متن کامل